Biochemical characterization of recombinant UDP-glucose: Sterol 3-O-Glycosyltransferase from Micromonospora rhodorangea ATCC 31603 and enzymatic biosynthesis of sterol-3-O-β-glucosides

Nguyen Huu Hoang, Sung Yong Hong, Nguyen Lan Huong, Je Won Park

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

A uridine diphosphate-glucose:sterol glycosyltransferase-encoding gene was isolated and cloned from the established fosmid library of Micromonospora rhodorangea ATCC 27932 that usually produces the aminoglycoside antibiotic geneticin. The gene consists of 1,185 base pairs and encodes a 41.4 kDa protein, which was heterologously expressed in Escherichia coli BL21(DE3). In silico analyses of the deduced gene product suggested that it is a member of the family 1 glycosyltransferases. The recombinant protein MrSGT was able to catalyze the transfer of a glucosyl moiety onto the C-3 hydroxy function in sterols (β-sitosterol, campesterol, and cholesterol), resulting in the corresponding steryl glucosides (β-sitosterol-3- O-β-D-glucoside, campesterol-3-O-β-D-glucoside, and cholesterol-3-O-β-D-glucoside). This enzyme prefers phytosterols to cholesterol, and also shows substrate flexibility to some extent, in that it could recognize a number of acceptor substrates.

Original languageEnglish
Pages (from-to)477-482
Number of pages6
JournalJournal of microbiology and biotechnology
Volume26
Issue number3
DOIs
Publication statusPublished - 2015 Dec 8

Fingerprint

Micromonospora
Uridine Diphosphate Glucose
Glycosyltransferases
Glucosides
Sterols
Cholesterol
Genes
Phytosterols
Aminoglycosides
Recombinant Proteins
Base Pairing
Computer Simulation
Libraries
Escherichia coli
Anti-Bacterial Agents
Enzymes
Proteins
campesterol
gamma-sitosterol

Keywords

  • Micromonospora rhodorangea
  • Phytosterol-3-O-β-D-glucosides
  • UDP-glucose sterol glycosyltransferase

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

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title = "Biochemical characterization of recombinant UDP-glucose: Sterol 3-O-Glycosyltransferase from Micromonospora rhodorangea ATCC 31603 and enzymatic biosynthesis of sterol-3-O-β-glucosides",
abstract = "A uridine diphosphate-glucose:sterol glycosyltransferase-encoding gene was isolated and cloned from the established fosmid library of Micromonospora rhodorangea ATCC 27932 that usually produces the aminoglycoside antibiotic geneticin. The gene consists of 1,185 base pairs and encodes a 41.4 kDa protein, which was heterologously expressed in Escherichia coli BL21(DE3). In silico analyses of the deduced gene product suggested that it is a member of the family 1 glycosyltransferases. The recombinant protein MrSGT was able to catalyze the transfer of a glucosyl moiety onto the C-3 hydroxy function in sterols (β-sitosterol, campesterol, and cholesterol), resulting in the corresponding steryl glucosides (β-sitosterol-3- O-β-D-glucoside, campesterol-3-O-β-D-glucoside, and cholesterol-3-O-β-D-glucoside). This enzyme prefers phytosterols to cholesterol, and also shows substrate flexibility to some extent, in that it could recognize a number of acceptor substrates.",
keywords = "Micromonospora rhodorangea, Phytosterol-3-O-β-D-glucosides, UDP-glucose sterol glycosyltransferase",
author = "Hoang, {Nguyen Huu} and Hong, {Sung Yong} and Huong, {Nguyen Lan} and Park, {Je Won}",
year = "2015",
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T1 - Biochemical characterization of recombinant UDP-glucose

T2 - Sterol 3-O-Glycosyltransferase from Micromonospora rhodorangea ATCC 31603 and enzymatic biosynthesis of sterol-3-O-β-glucosides

AU - Hoang, Nguyen Huu

AU - Hong, Sung Yong

AU - Huong, Nguyen Lan

AU - Park, Je Won

PY - 2015/12/8

Y1 - 2015/12/8

N2 - A uridine diphosphate-glucose:sterol glycosyltransferase-encoding gene was isolated and cloned from the established fosmid library of Micromonospora rhodorangea ATCC 27932 that usually produces the aminoglycoside antibiotic geneticin. The gene consists of 1,185 base pairs and encodes a 41.4 kDa protein, which was heterologously expressed in Escherichia coli BL21(DE3). In silico analyses of the deduced gene product suggested that it is a member of the family 1 glycosyltransferases. The recombinant protein MrSGT was able to catalyze the transfer of a glucosyl moiety onto the C-3 hydroxy function in sterols (β-sitosterol, campesterol, and cholesterol), resulting in the corresponding steryl glucosides (β-sitosterol-3- O-β-D-glucoside, campesterol-3-O-β-D-glucoside, and cholesterol-3-O-β-D-glucoside). This enzyme prefers phytosterols to cholesterol, and also shows substrate flexibility to some extent, in that it could recognize a number of acceptor substrates.

AB - A uridine diphosphate-glucose:sterol glycosyltransferase-encoding gene was isolated and cloned from the established fosmid library of Micromonospora rhodorangea ATCC 27932 that usually produces the aminoglycoside antibiotic geneticin. The gene consists of 1,185 base pairs and encodes a 41.4 kDa protein, which was heterologously expressed in Escherichia coli BL21(DE3). In silico analyses of the deduced gene product suggested that it is a member of the family 1 glycosyltransferases. The recombinant protein MrSGT was able to catalyze the transfer of a glucosyl moiety onto the C-3 hydroxy function in sterols (β-sitosterol, campesterol, and cholesterol), resulting in the corresponding steryl glucosides (β-sitosterol-3- O-β-D-glucoside, campesterol-3-O-β-D-glucoside, and cholesterol-3-O-β-D-glucoside). This enzyme prefers phytosterols to cholesterol, and also shows substrate flexibility to some extent, in that it could recognize a number of acceptor substrates.

KW - Micromonospora rhodorangea

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KW - UDP-glucose sterol glycosyltransferase

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